The present invention relates to proteins which lack tyrosine kinase activity and dimerize with members of the erbB family of receptors; to nucleic acid molecules that encode such proteins; to pharmaceutical compositions that comprise such nucleic acid molecules in combination with delivery vehicles which facilitate transfer of the nucleic acid molecule to a cell; and to methods of preventing tumors and treating individuals having tumors by administering such pharmaceutical compositions. The present invention relates to compositions which are useful to convert tumor cells that are resistant to radiation- and/or chemical-induced cell death into cells which are sensitive to radiation. The present invention relates to methods of treating individuals who have tumors by administering such compositions in combination with radiation and/or chemotherapy.
The erbB family of receptors includes erbB1 (EGFR), erbB2 (p185), erbB3 and erbB4. Ullrich, et al. (1984) Nature 309, 418-425, which is incorporated herein by reference, describes EGFR. Schechter, A. L., et al. (1984) Nature 312, 513-516, and Yamamoto, T., et al. (1986) Nature 319, 230-234, which are each incorporated herein by reference, describe p185neu/erbB2. Kraus, M. H., et al. (1989) Proc. Natl. Acad. Sci. USA 86, 9193-9197 which is incorporated herein by reference, describes erbB3. Plowman, G. D., (1993) Proc. Natl. Acad. Sci. USA 90, 1746-1750, which is incorporated herein by reference, describes erbB4.
The rat cellular protooncogene c-neu and its human counterpart c-erbB2 encode 185 kDa transmembrane glycoproteins termed p185. Tyrosine kinase (tk) activity has been linked to expression of the transforming phenotype of oncogenic p185 (Bargmann et al., Proc. Natl. Acad. Sci. USA, 1988, 85, 5394; and Stern et al., Mol. Cell. Biol., 1988, 8, 3969, each of which is incorporated herein by reference). Oncogenic neu was initially identified in rat neuroglioblastomas (Schechter et al., Nature, 1984, 312, 513, which is incorporated herein by reference) and was found to be activated by a carcinogen-induced point mutation generating a single amino acid substitution, a Val to Glu substitution at position 664, in the transmembrane region of the transforming protein (Bargmann et al., Cell, 1986, 45, 649, which is incorporated herein by reference). This alteration results in constitutive activity of its intrinsic kinase and in malignant transformation of cells (Bargmann et al., EMBO J, 1988, 7,2043, which is incorporated herein by reference). The activation of the oncogenic p185 protein tyrosine kinase appears to be related to a shift in the molecular equilibrium from monomeric to dimeric forms (Weiner et al., Nature, 1989, 339, 230, which is incorporated herein by reference).
Overexpression of c-neu or c-erbB2 to levels 100-fold higher than normal (i.e., greater than 106 receptors/cell) also results in the transformation of NIH3T3 cells (Chazin et al., Oncogene, 1992, 7, 1859; DiFiore et al., Science, 1987, 237, 178; and DiMarco et al., Mol. Cell. Biol., 1990, 10, 3247, each of which is incorporated herein by reference). However, NIH3T3 cells or NR6 cells which express cellular p185 at the level of 105 receptors/cell are not transformed (Hung et al., Proc. Natl. Acad. Sci. USA, 1989, 86, 2545; and Kokai et al., Cell, 1989, 58, 287, each of which is incorporated herein by reference), unless co-expressed with epidermal growth factor receptor (EGFR), a homologous tyrosine kinase (Kokai et al., Cell, 1989, 58, 287, which is incorporated herein by reference). Thus, cellular p185 and oncogenic p185 may both result in the transformation of cells.
Cellular p185 is highly homologous with EGFR (Schechter et al., Nature, 1984, 312, 513; and Yamamoto et al., Nature, 1986, 319, 230, each of which is incorporated herein by reference) but nonetheless is distinct. Numerous studies indicate that EGFR and cellular p185 are able to interact (Stern et al., Mol. Cell. Biol., 1988, 8, 3969; King et al., EMBO J., 1988, 7, 1647; Kokai et al., Proc. Natl. Acad. Sci. USA, 1988, 85, 53 89; and Dougall et al., J. Cell. Biochem., 1993, 53, 61; each of which is incorporated herein by reference). The intermolecular association of EGFR and cellular p185 appear to up-regulate EGFR function (Wada et al., Cell, 1990, 61, 1339, which is incorporated herein by reference). In addition, heterodimers which form active kinase complexes both in vivo and in vitro can be detected (Qian et al., Proc. Natl. Acad. Sci. USA, 1992, 89, 1330, which is incorporated herein by reference).
Similarly, p185 interactions with other erbB family members have been reported (Carraway et al., Cell 1994, 78, 5-8; Alroy et al., FEBS Lett. 1997, 410, 83-86; Riese et al., Mol. Cell. Biol. 1995, 15, 5770-5776; Tzahar et al., EMBO J. 1997, 16, 4938-4950; Surden et al., Neuron 1997, 18, 847-855; Pinkas-Kramarski et al., Oncogene 1997, 15, 2803-2815; each of which is incorporated herein by reference). Human p185 forms heterodimers with either erbB3 or erbB4 under physiologic conditions, primarily in cardiac muscle and the nervous system, particularly in development.
Cellular p185 proteins are found in adult secretory epithelial cells of the lung, salivary gland, breast, pancreas, ovary, gastrointestinal tract, and skin (Kokai et al., Proc. Natl. Acad. Sci. USA, 1987, 84, 8498; Mori et al., Lab. Invest., 1989, 61, 93; and Press et al., Oncogene, 1990, 5, 953; each of which is incorporated herein by reference). Recent studies have found that the amplification of c-erbB2 occurs with high frequency in a number of human adenocarcinomas such as gastric (Akiyama et al., Science, 1986, 232, 1644, which is incorporated herein by reference), lung (Kern et al., Cancer Res., 1990, 50, 5184, which is incorporated herein by reference) and pancreatic adenocarcinomas (Williams et al., Pathobiol., 1991, 59, 46, which is incorporated herein by reference). It has also been reported that increased c-erbB2 expression in a subset of breast and ovarian carcinomas is linked to a less optimistic clinical prognosis (Slamon et al., Science, 1987, 235, 177; and Slamon et al., Science, 1989, 244, 707, each of which is incorporated herein by reference). Heterodimeric association of EGFR and p185 has also been detected in human breast cancer cell lines, such as SK-Br-3 (Goldman et al., Biochemistry, 1990, 29, 11024, which is incorporated herein by reference), and transfected cells (Spivak-Kroizman et al., J Biol. Chem., 1992, 267, 8056, which is incorporated herein by reference). Additionally, cases of erbB2 and EGFR coexpression in cancers of the breast and prostate have been reported. In addition, heterodimeric association of p185 and erbB3 as well as heterodimeric association of p185 and erbB4 have also been detected in human cancers. Coexpression of erbB2 and erbB3 has been observed in human breast cancers. Coexpression of EGFR, erbB2, and erbB3 has been seen in prostate carcinoma.
Amplification and/or alteration of the EGFr gene is frequently observed in glial tumor progression (Sugawa, et al. (1990) Proc. Natl. Acad. Sci. 87: 8602-8606; Ekstrand, et al. (1992) Proc. Natl. Acad. Sci. 89: 4309-4313), particularly in glioblastoma, the most malignant glial tumor (Libermann, et al Supra; Wong, et al. Supra; James, et al. (1988) Cancer Res. 48: 5546-5551; Cavenee, W. K. (1992) Cancer 70: 1788-93; Nishikawa, et al., (1994) Proc. Natl. Acad. Sci. 91: 7727-7731; Schlegel, et al. (1994) Int J. Cancer 56: 72-77). A significant proportion of these tumors show EGFr amplification with or without gene alteration (Ekstrand, et al Supra; Libermann, et al. Supra; Wong, et al. (1987) Proc. Natl. Acad. Sci. 84:6899-6903), and this has been correlated with a shorter interval to disease recurrence and poorer survival (Schlegel, et al. Supra).
EGFr amplification can be associated with aberrant EGFr transcripts along with normal EGFr transcripts (Sugawa, et al Supra). Frequent amplification and subsequent structural alteration suggests the EGFr may be important for the maintenance of the phenotype of malignant glioma. A frequently observed EGFr mutant has been identified in a subset of human glioblastomas and results from an in-frame truncation of 801 bp (corresponding to exons 2-7) in the extracellular domain of the receptor (Sugawa, et al. Supra; Ekstrand, et al Supra; Malden, et al. (1988) Cancer Res. 48: 2711-2714; Humphrey, et al. (1990) Proc. Natl. Acad. Sci. 87: 4207-4211; Wong, et al. (1992) Proc. Natl. Acad. Sci. 89: 2965-2969), which is thought to result in constitutive kinase activation and may also affect the ligand-binding properties of the molecule (Nishikawa, et al. Supra; Callaghan, et al. (1993) Oncogene 8: 2939-2948).
Observed mutations of EGFr in human epithelial malignancies consist of overexpression with or without amplification and, less commonly, of coding sequence alterations. Oncogenic transformation caused by mutants of EGFr appear to be tissue-specific and have been observed in erythroid leukemia, fibrosarcoma, angiosarcoma, melanoma, as well as glioblastoma (Carter, et al. (1994) Crit Rev Oncogenesis 5: 389-428). Overexpression of the normal EGFr may cause oncogenic transformation in certain cases, probably in an EGF-dependent manner (Carter, et al. Supra; Haley, et al. (1989) Oncogene 4: 273-283). Transfection of high amounts of wild-type EGFr into NIH3T3 cells results in ligand-dependent but incomplete transformation (Yamazaki, et al. (1990) Jpn. J. Cancer Res. 81: 773-779). Overexpression may cause altered cell-cycle regulation of the EGFr kinase, and contribute to the transformed state, as has been observed for oncogenic p185neu (Kiyokawa, et al. (1995) Proc. Natl. Acad. Sci. 92:1092-1096).
There is a need for therapeutic compositions useful to treat individuals identified as having erbB-mediated tumors. There is a need to develop prophylactic compositions for individuals susceptible to developing erbB-mediated tumors. There is a need for methods of treating individuals identified as having erbB-mediated tumors. There is a need to methods of preventing individuals who are susceptible to developing erbB-mediated tumors from developing such tumors.
The present invention relates to nucleic acid molecules which comprise a nucleotide sequence that encodes a protein that lacks tyrosine kinase activity and dimerizes with members of the erbB family of receptors, such as erbB1 (EGFR), erbB2 (p185), erbB3 and/or erbB4.
The present invention relates to nucleic acid molecules which comprise a nucleotide sequence that encodes a protein that lacks tyrosine kinase activity and dimerizes with human EGFR and human 185.
The present invention relates to nucleic acid molecules in combination with delivery components in which the nucleic acid molecules comprise a nucleotide sequence that encodes a protein that lacks tyrosine kinase activity and dimerizes with members of the erbB family of receptors, such as erbB1 (EGFR), erbB2 (p185), erbB3 and/or erbB4.
The present invention relates to nucleic acid molecules in combination with delivery components in which the nucleic acid molecules comprise a nucleotide sequence that encodes a protein that lacks tyrosine kinase activity and dimerizes with human EGFR and human p185.
The present invention relates to recombinant viral vectors which comprise nucleic acid molecules that include a nucleotide sequence that encodes a protein that lacks tyrosine kinase activity and dimerizes with members of the erbB family of receptors, such as erbB1 (EGFR), erbB2 (p185), erbB3 and/or erbB4.
The present invention relates to recombinant viral vectors which comprise nucleic acid molecules that include a nucleotide sequence that encodes a protein that lacks tyrosine kinase activity and dimerizes with human EGFR and human p185.
The present invention relates to a pharmaceutical composition comprising a nucleic acid molecule in combination with delivery components. The nucleotide sequence of the nucleic acid molecule encodes a protein that lacks tyrosine kinase activity and dimerizes with members of the erbB family of receptors, such as erbB1 (EGFR), erbB2 (p185), erbB3 and/or erbB4, and preferably a protein that lacks tyrosine kinase activity and dimerizes with human EGFR and human p185.
The present invention relates to a pharmaceutical composition comprising recombinant viral vectors that include nucleic acid molecules with a nucleotide sequence that encodes a protein that lacks tyrosine kinase activity and dimerizes with human EGFR and human p185.
The present invention relates to a method of treating an individual identified as undergoing erbB-mediated cellular transformation. The treatment includes administering to the individual a pharmaceutical composition comprising a nucleic acid molecule in combination with delivery components in an amount sufficient to reverse the cellular transformation. The nucleic acid sequence encodes a protein that lacks tyrosine kinase activity and dimerizes with members of the erbB family of receptors, such as erbB1 (EGFR), erbB2 (p185), erbB3 and/or erbB4. The delivery components may be viral particles and the nucleic acid molecule may be a viral genome.
The present invention relates to a method of treating an individual identified as undergoing p185-mediated cellular transformation. The treatment includes administering to the individual a pharmaceutical composition comprising a nucleic acid molecule in combination with delivery components in an amount sufficient to reverse the cellular transformation. The nucleic acid sequence encodes a protein that lacks tyrosine kinase activity and dimerizes with human EGFR and human p185. The delivery components may be viral particles and the nucleic acid molecule may be a viral genome.
The present invention relates to methods of preventing erbB-mediated cellular transformation in an individual identified as susceptible to erbB-mediated cellular transformation. The methods include administering to the individual a pharmaceutical composition comprising a nucleic acid molecule in combination with delivery components in an amount sufficient to prevent the cellular transformation. The nucleic acid sequence encodes a protein that lacks tyrosine kinase activity and dimerizes with members of the erbB family of receptors, such as erbB1 (EGFR), erbB2 (p185), erbB3 and/or erbB4. The delivery components may be viral particles and the nucleic acid molecule may be a viral genome.
The present invention relates to methods of preventing p185-mediated cellular transformation in an individual identified as being susceptible to p185-mediated cellular transformation. The methods include administering to the individual a pharmaceutical composition comprising a nucleic acid molecule in combination with delivery components in an amount sufficient to prevent the cellular transformation. The nucleic acid sequence encodes a protein that lacks tyrosine kinase activity and dimerizes with human EGFR and human p185. The delivery components may be viral particles and the nucleic acid molecule may be a viral genome.
The present invention relates to methods treating individuals who have erbB protein mediated tumors comprising the steps of administering to such individuals, nucleic acid molecules that encode a protein that dimerizes with said erbB protein and that is deficient in tyrosine kinase activity, and exposing said individual to a therapeutically effective amount of anti-cancer radiation and/or administering to said individual a therapeutically effective amount of an anti-cancer chemotherapeutic.
The present invention relates to methods of treating individuals who have erbB protein mediated tumors comprising the steps of first administering to the individuals a composition which inhibits formation of erbB protein dimers that produce elevated tyrosine kinase activity in a tumor cell, followed by exposing the individuals to a therapeutically effective amount of anti-cancer radiation.
The present invention relates to methods of treating an individual who have tumors that are characterized by tumor cells that have multimeric receptor ensembles which provide kinase activity associated with a transformed phenotype. The methods comprise the steps of administering to the individual, a composition that disrupts the kinase activity associated with the multimeric receptor ensemble; and exposing the individual to a therapeutic amount of gamma radiation.
The present invention relates to methods of treating individuals who have tumors that are characterized by tumor cells that have multimeric receptor ensembles which provide kinase activity associated with a transformed phenotype. The methods comprise the steps of administering to the individual, a active agent which is not an antibody, such as a peptide, non-proteinaceous compound or nucleic acid molecules that encodes a protein that disrupts the kinase activity associated with the multimeric receptor ensemble; and exposing the individual to a therapeutic amount of gamma radiation and/or administering a therapeutic amount of a cytotoxic chemotherapeutic agent to the individual.
The present invention relates to methods of treating individuals who have tumors that are characterized by tumor cells that comprise an EGFR species such as wild type or mutant EGFR, The method comprises the steps of administering to the individual, a composition that disrupts kinase activity mediated by an EGFR species; and exposing the individual to a therapeutic amount of gamma radiation and/or administering a therapeutic amount of a cytotoxic chemotherapeutic agent to said individual.
The present invention relates to methods of treating an individual who has an erbB protein mediated tumor comprising the steps of administering to the individual a nucleic acid molecule that encodes a protein that dimerizes with the erbB protein and that is deficient in tyrosine kinase activity, and exposing the individual to a therapeutically effective amount of anti-cancer radiation and/or administering to the individual a therapeutically effective amount of an anti-cancer chemotherapeutic. In some embodiments, the erbB-protein mediated tumor is a p185-mediated tumor. In some embodiments, the erbB-protein mediated tumor is an EGFR-mediated tumor. In some embodiments, the erbB-protein mediated tumor is a glial tumor. In some embodiments, the erbB-protein mediated tumor is a glioblastoma. In some embodiments, the administration of the nucleic acid molecule is by intratumor administration. In some embodiments, the individual has surgery prior to administration of the nucleic acid molecule. In some embodiments, the protein comprises a p185 ectodomain. In some embodiments, the protein comprises a rat neu transmembrane region with a val to glu mutation at amino acid 664. In some embodiments, the nucleic acid molecule is the viral genome of a recombinant adenovirus. In some embodiments, the nucleic acid molecule comprises a coding sequences operably linked to regulatory elements for translation in cells of the individual, the coding sequence comprises: a truncated rat neu gene with a stop codon at amino acid 691; a truncated rat neu gene with a stop codon at amino acid 691 and valxe2x86x92glu mutation at amino acid 664; a chimeric p185 gene comprising human p185c-erbB2 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691; a chimeric p185 gene comprising human p185c-erbB2 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691 and a valxe2x86x92glu mutation at amino acid 664; a chimeric p185 gene comprising human EGFR ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691; a chimeric p185 gene comprising human EGFR ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691 and a valxe2x86x92glu mutation at amino acid 664; a chimeric p185 gene comprising human erbB3 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691; a chimeric p185 gene comprising human erbB3 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691 and a valxe2x86x92glu mutation at amino acid 664; a chimeric p185 gene comprising human erbB4 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691; or a chimeric p185 gene comprising human erbB4 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691 and a valxe2x86x92glu mutation at amino acid 664. In some embodiments, the individual is exposed to a therapeutically effective amount of anti-cancer radiation. In some embodiments, the individual is administered a therapeutically effective amount of anti-cancer chemotherapeutic.
The present invention relates to methods of treating an individual who has an erbB protein mediated brain tumor comprising the step administering to the individual a nucleic acid molecule that encodes a protein that dimerizes with the erbB protein and is deficient in tyrosine kinase activity. In some embodiments, the protein comprises a p185 ectodomain. In some embodiments, the nucleic acid molecule contains a rat neu transmembrane region with a val to glu mutation at amino acid 664. In some embodiments, the erbB-protein mediated tumor is an EGFR-mediated tumor. In some embodiments, the erbB-protein mediated tumor is a mutant EGFR-mediated tumor. In some embodiments, the erbB-protein mediated tumor is a glioblastoma. In some embodiments, the individual is exposed to a therapeutically effective amount of anti-cancer radiation and/or administered a therapeutically effective amount of an anti-cancer chemotherapeutic. In some embodiments, the administration of the nucleic acid molecule is by direct injection into the tumor. In some embodiments, the administration of the nucleic acid molecule is by direct injection into the tumor using stereotaxic surgical procedures. In some embodiments, the individual has surgery to remove bulk tumor prior to administration of the nucleic acid molecule. In some embodiments, the nucleic acid molecule is the viral genome of a recombinant adenovirus. In some embodiments, the nucleic acid molecule comprises a coding sequences operably linked to regulatory elements for translation in cells of the individual, and the coding sequence comprises: a truncated rat neu gene with a stop codon at amino acid 691 (N691stop construct); a truncated rat neu gene with a stop codon at amino acid 691 and a valxe2x86x92glu mutation at amino acid 664; a chimeric p185 gene comprising human p185c-erbB2 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691; a chimeric p185 gene comprising human p185c-erbB2 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691 and a valxe2x86x92glu mutation at amino acid 664; a chimeric p185 gene comprising human EGFR ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691; a chimeric p185 gene comprising human EGFR ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691 and a valxe2x86x92glu mutation at amino acid 664; a chimeric p185 gene comprising human erbB3 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691; a chimeric p185 gene comprising human erbB3 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691 and a valxe2x86x92glu mutation at amino acid 664; a chimeric p185 gene comprising human erbB4 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691; or a chimeric p185 gene comprising human erbB4 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691 and a valxe2x86x92glu mutation at amino acid 664.
The present invention relates to methods of inhibiting proliferation of a mutant EGFR-mediated tumor cell comprising the step of delivering to the cell a nucleic acid molecule that encodes a protein that dimerizes with mutant EGFR and is deficient in tyrosine kinase activity. In some embodiments, the protein comprises a p185 ectodomain. In some embodiments, the protein contains a rat neu transmembrane region with a val to glu mutation at amino acid 664. In some embodiments, the mutant EGFR-mediated tumor cell is a glioblastoma cell.
The present invention relates to methods of treating an individual who has an erbB protein mediated tumor comprising the steps of administering to the individual a composition which inhibits formation of erbB protein dimers that produce elevated tyrosine kinase activity in a tumor cell, and exposing the individual to a therapeutically effective amount of anti-cancer radiation. In some embodiments, the erbB-protein mediated tumor is a p185-mediated tumor. In some embodiments, the erbB-protein mediated tumor is an EGFR-mediated tumor. In some embodiments, the erbB-protein mediated tumor is a glial tumor. In some embodiments, the erbB-protein mediated tumor is a glioblastoma. In some embodiments, the administration of the composition is by intratumor administration. In some embodiments, the individual has surgery prior to administration of the composition. In some embodiments, the composition that is administered to a patient comprises a compound that interacts with an erbB protein in a tumor cell to alter the erbB protein sufficient to result in a decreased propensity of it to dimerize with another erbB protein. In some embodiments, the compound that interacts with an erbB protein in a tumor cell to alter the erbB protein sufficient to result in a decreased propensity of it to dimerize with another erbB protein is an antibody. In some embodiments, the antibody is a monoclonal antibody. In some embodiments, the composition that is administered to a patient comprises a compound that competitively interacts with an erbB protein in a tumor cell to competitively inhibit dimer formation with another erbB protein and prevents elevated tyrosine kinase activity. In some embodiments the compound that competitively interacts with an erbB protein in a tumor cell to competitively inhibit dimer formation with another erbB protein is a peptide. In some embodiments, the compound that competitively interacts with an erbB protein in a tumor cell to competitively inhibit dimer formation with another erbB protein is an antibody. In some embodiments, the composition that is administered to the tumor cell is a nucleic acid molecule that encodes a protein that competitively interacts with an erbB protein in a tumor cell to competitively inhibit dimer formation with another erbB protein. In some embodiments, the protein is a mutant or truncated kinase deficient erbB protein. In some embodiments, the protein is a mutant or truncated kinase deficient p185 protein. In some embodiments, the protein interacts with the transmembrane region of the one erbB protein. In some embodiments, the protein comprises a rat neu transmembrane region with a val to glu mutation at amino acid 664. In some embodiments, the protein interacts with the ectodomain region of the one erbB protein. In some embodiments, the protein comprises a p185 ectodomain. In some embodiments, the nucleic acid molecule is administered by intratumor administration. In some embodiments, the individual has surgery prior to administration of the nucleic acid molecule. In some embodiments, the nucleic acid molecule is the viral genome of a recombinant adenovirus. In some embodiments, the nucleic acid molecule comprises a coding sequences operably linked to regulatory elements for translation in cells of the individual, and the coding sequence comprises: a truncated rat neu gene with a stop codon at amino acid 691; a truncated rat neu gene with a stop codon at amino acid 691 and a valxe2x86x92glu mutation at amino acid 664; a chimeric p185 gene comprising human p185c-erbB2 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691; a chimeric p185 gene comprising human p185c-erbB2 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691 and a valxe2x86x92glu mutation at amino acid 664; a chimeric p185 gene comprising human EGFR ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691; a chimeric p185 gene comprising human EGFR ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691 and a valxe2x86x92glu mutation at amino acid 664; a chimeric p185 gene comprising human erbB3 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691; a chimeric p185 gene comprising human erbB3 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691 and a valxe2x86x92glu mutation at amino acid 664; a chimeric p185 gene comprising human erbB4 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691; or a chimeric p185 gene comprising human erbB4 ectodomain linked to rat neu transmembrane with a stop codon at amino acid 691 and a valxe2x86x92glu mutation at amino acid 664.
The present invention relates to methods of treating an individual who has a tumor, wherein the tumor is characterized by tumor cells that have multimeric receptor ensembles which provide kinase activity associated with a transformed phenotype. The method comprises the steps of administering to the individual, a composition that disrupts the kinase activity associated with the multimeric receptor ensemble; and exposing the individual to a therapeutic amount of gamma radiation. In some embodiments, the tumor is characterized by tumor cells that have multimeric receptor ensembles selected from the group consisting of: erbB homodimers, erbB heterodimers, and multimers of platelet derived growth factor receptors. In some embodiments, the tumor is characterized by tumor cells that have erbB homodimers that are mutant EGFR homodimers or p185 homodimers. In some embodiments, the tumor is characterized by tumor cells that have erbB heterodimers that are p185/EGFR heterodimers, p185/mutant EGFR heterodimers, p185/erbB3 heterodimers; p185/erbB4 heterodimers or EGFR/mutant EGFR heterodimers. In some embodiments, the composition that disrupts the kinase activity associated with the multimeric receptor ensemble comprises an active agent selected from the group consisting of antibodies, peptides, and non-proteinaceous kinase inhibitors. In some embodiments, the composition that disrupts the kinase activity associated with the multimeric receptor ensemble comprises an active agent that is a nucleic acid molecule that encodes a protein or peptide which interacts with a monomeric component of the ensemble to prevent the monomeric component from interacting with a second monomeric component of the ensemble.
The present invention relates to methods of treating an individual who has a tumor, wherein the tumor is characterized by tumor cells that have multimeric receptor ensembles which provide kinase activity associated with a transformed phenotype. The method comprises the steps of administering to the individual, a small peptide, non-proteinaceous compound or nucleic acid molecules that encodes a non-antibody protein or peptide that disrupts the kinase activity associated with the multimeric receptor ensemble; and exposing the individual to a therapeutic amount of gamma radiation and/or administering a therapeutic amount of a cytotoxic chemotherapeutic agent to the individual. In some embodiments, the tumor is characterized by tumor cells that have multimeric receptor ensembles selected from the group consisting of: erbB homodimers, erbB heterodimers, and multimers of platelet derived growth factor receptors. In some embodiments, the tumor is characterized by tumor cells that have erbB homodimers that are mutant EGFR homodimers or p185 homodimers. In some embodiments, the tumor is characterized by tumor cells that have erbB heterodimers that are p185/EGFR heterodimers, p185/mutant EGFR heterodimers, p185/erbB3 heterodimers; p185/erbB4 heterodimers or EGFR/mutant EGFR heterodimers. In some embodiments, the composition that disrupts the kinase activity associated with the multimeric receptor ensemble comprises an active agent selected from the group consisting of peptides, and non-proteinaceous kinase inhibitors. In some embodiments, the composition that disrupts the kinase activity associated with the multimeric receptor ensemble comprises an active agent that is a nucleic acid molecule that encodes a protein or peptide which interacts with a monomeric component of the ensemble to prevent the monomeric component from interacting with a second monomeric component of the ensemble.
The present invention relates to methods of treating an individual who has a tumor, wherein the tumor is characterized by tumor cells that comprise an EGFR species. The methods comprise the steps of administering to the individual, a composition that disrupts kinase activity mediated by an EGFR species; and exposing the individual to a therapeutic amount of gamma radiation and/or administering a therapeutic amount of a cytotoxic chemotherapeutic agent to the individual. In some embodiments, the EGFR species is a mutant EGFR. In some embodiments, the composition that disrupts kinase activity mediated by an EGFR species comprises an active agent selected from the group consisting of antibodies, peptides, and non-proteinaceous kinase inhibitors. In some embodiments, the composition that disrupts kinase activity mediated by an EGFR species comprises an active agent that is a nucleic acid molecule that encodes a protein or peptide which interacts with a molecule of an EGFR species to prevent the molecule or from forming a kinase-activity elevating multimeric ensemble with a second molecule. In some embodiments, the individual is administered a cytotoxic chemotherapeutic agent.